Tentative identification of in vitro metabolites of O‐acetylpsilocin (psilacetin, 4‐AcO‐DMT) by UHPLC‐Q‐Orbitrap MS
Drug Testing and Analysis – March 21, 2022
Source: OpenAlex
Summary
The psychedelic tryptamine 4-AcO-DMT undergoes significant drug metabolism, yielding 15 distinct metabolites. Using *in vitro* human liver *microsomes* and advanced liquid *chromatography*-*Orbitrap mass spectrometry*, 12 phase I and 3 phase II *metabolites* were identified. Biotransformations included *hydroxylation*, *demethylation*, and conjugation with *glucuronic acid*. The hydrolysis *metabolite* was most abundant. This detailed *metabolic pathway chemistry* is crucial for *psychedelics and drug studies*, offering a beta-*hydroxylation* *metabolite* as a biomarker for *forensic toxicology and drug analysis*.
Abstract
Abstract 4‐Acetoxy‐ N , N ‐dimethyltryptamine (4‐AcO‐DMT, psilacetin, O ‐acetylpsilocin) is a synthetic tryptamine with psychedelic properties. Psilacetin may also act as precursor drug of psilocin, similar to psilocybin, but little is known about its metabolism. In this study, the phase I and phase II in vitro metabolism of 4‐AcO‐DMT was investigated with pooled human liver microsomes, and the reaction mixture was analyzed using liquid chromatography‐quadrupole/electrostatic field orbitrap mass spectrometry. Fifteen metabolites were formed after incubation of pooled human liver microsomes with 4‐AcO‐DMT (12 phase I metabolites and 3 phase II metabolites). The proposed metabolite structures were based on accurate mass analysis and MS/MS fragmentation patterns. The biotransformations included hydrolysis, hydroxylation, N ‐demethylation, oxidation, and conjugation with glucuronic acid. The hydrolysis metabolite was the most abundant compound. For the development of new methods for the identification of 4‐AcO‐DMT consumption, the beta‐hydroxylation metabolite of 4‐AcO‐DMT (M2‐1) is recommended as a biomarker. The data reported in this work might be applicable to metabolic transformation of 4‐AcO‐DMT in vivo and also forensically helpful.